Search

Background. Mycotoxins are toxic secondary metabolites, which are synthesized from a large number of intermediates from the primary metabolism of saprophytic molds. Aflatoxins, due to their genotoxic and carcinogenic effects, are the most important group of mycotoxins from the aspect of their impact on human and animal health. Of all known aflatoxins, AFB1 is the most frequent, with the most harmful impact on human and animal health. Scope and Approach. Due to their prevalence and toxicity, monitoring the presence of aflatoxins in the food chain is required. The scope of this paper is to provide information on the presence of aflatoxins in animal feed and in milk. This paper describes temperature increases in Europe that are contributing to the increased presence of aflatoxins in food, as well as aflatoxin prevention and protection measures. Key Findings and Conclusions. During the last decade, serious contamination of corn with aflatoxins was recorded in southern Europe. In the summer of 2012, Serbia recorded high concentrations of aflatoxins in corn and milk. Based on climate change data, it is expected that aflatoxin contamination in corn will become more frequent. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 31034]

Background/Aim. Factors determining the risk of Lyme disease (LD) may be followed in terms of the type of habitat. The evaluation of the risk of Borrelia burgdorferi (B. burgdorferi) transmission in humans on Ixodes ricinus (I. ricinus) tick habitats is done by means of the ecological risk index, which determines the tick habitat, abundance and infection rate. The aim of this paper was to determine the value of ecological risk index (potential and actual risk – PR and AR) on green areas in 9 pаrks in Belgrade and establish the correlation of this index with tick bites in humans. Methods. Ticks were collected in parks by means of the flag hour method and examined for the presence of LD cause in dark-field microscopic analysis. Point values were assigned to certain parameters and potential and actual risk index evaluated for each habitat. The data on tick bites from the surveyed habitats were obtained from the Protocol of patients bitten by ticks of the Sector for Preventive Medicine, Institute of Epidemiology, Military Medical Academy in Belgrade. Analysis of variance (ANOVA), Tukey test and Pearson ´s coefficient were used in statistical analysis of data. Results. In Belgrade´s park habitats a high PR of B. burgdorferi transmission was determined, while AR for 4 habitats was categorised as PR, and limited for other 5 habitats. Statistically, in terms of AR values the following habitats were significantly different (p < 0.05): Hаjd Pаrk and Tаšmаjdаn; Hаjd Pаrk and Kаlemegdаn; Hаjd Pаrk and Pionirski Pаrk; Hаjd Pаrk and Bаnovo Brdo; Topčider and Tаšmаjdаn; Topčider and Kаlemegdаn; Topčider and Pionirski Pаrk; Topčider and Bаnovo Brdo Park. A statistically significant correlation (p < 0.05) between bites of adults and the number of bites of infected adults with the AR value was established. Conclusion. In parks of Belgrade, there is a limited AR of B. burgdorferi transmission on the average. The AR values vary from limited to the potential, depending on the ecological features of habitat, the number of collected ticks and their infection rate. In view of the correlation of AR with the bites in humans, this index is significant for assessing LD risk.

Background/Aim. The first case of human Lyme borreliosis (LB) in Serbia was recorded in 1987. The number of reported LB cases has increased in the past decade. The aim of this study was to estimate the density of Ixodes ricinus (I. ricinus) ticks, the prevalence of Borrelia burgdorferi sensu lato (B. burgdorferi) in them, and entomological risk index (ERI) at 19 Belgrade localities which were grouped into three categories (forests, parkforests, parks). The values of ERI were compared with the number of tick bites in humans. Methods. Ticks were collected monthly by using the flag hours method and the infection rate was determined by using dark field microscopy. The ERI value was calculated for each locality where the ticks were collected. The related data about tick bites was obtained from the patient protocol of the Institute of Epidemiology, Military Medical Academy, Belgrade. Results. The total number of collected ticks, the number of nymphs and the infection rates of the nymphs were significantly higher in forests (p < 0.05) than park-forests and parks. Statistically, the ERI value was significantly higher in forests than parks of Belgrade (χ2 = 7.78, p < 0.01). In March and July, the ERI value was also significantly higher in forests, than park-forests (p < 0.01) and parks (p < 0.01). May was the month with the highest ERI value in each ecological category (forests p < 0.05; park-forests p < 0.01; parks p < 0.001). However, the number of tick bites in humans did not correlate with ERI values. Conclusion. The obtained results indicate that the risk of tick bite and human exposure to B. burgdorferi sensu lato is present at all selected localities in Belgrade. For a more comprehensive Lyme disease risk assessment the method of entomological risk index assessment should be combined with other methods, taking into consideration all tick stages and the behaviour and habits of people who may get infected B. burgdorferi sensu lato.

Phenothiazines are a large group of heterocyclic, aromatic molecules with nitrogen and sulphur between two benzene rings. Their derivatives, N-alkylphenothiazines have supstitutient on heterocyclic nitrogen atom which gives them different properties. Also, a series of these molecules have substitution on carbon atom at place 2 of phenothiazine benzene ring. Alkylphenothiazines contain aminoalkyl substituent and their alkyl, acyl and sulphonil derivatives, as well as monocyclic and bicyclic heterocycles attached at thiazine nitrogen atom or directly linked to benzene ring. The N-alkylphenothiazines have been known as antipsychotic drugs, but they also possess antibacterial, antifungal, anticancer activity, and ability to react with macromolecules and to coordinate to the metals. Metal complexes with N-alkylphenothiazines are biological active compounds with different antimicrobial activities and cytotoxic effect against tumor cell lines. Their large field of application of N-alkylphenothiazines is very attractive in terms of synthesis of new related derivatives, metal complexes, studying their properties and applications. This article presents a review of the literature and a contemporary view at N-alkylphenothiazines - their synthesis and application, as well as their metal complexes which have promising biological effects. [Projekat Ministarstva nauke Republike Srbije, br. 172014]

In the territory of Belgrade, at 16 sites (15 of test and one control), which belong to different enviromental categories such as: parks (Topcider, Hajd park, park Banovo brdo, Sumice, Usce), park-forests (Kosutnjak, Zvezdara, SP Jajinci, Banjica, Ada Ciganlija) and forests (Avala, Lipovica, Bojcinska forest, Manastirska forest, Makis), there were investigated potential and actual risk of transmission of Borrelia burgdorferi sensu lato. Suplja Stena was taken as a control site, where no chemical treatment was performed. Once of month, from March to October 2012, ticks were collected by flag-hours (drag-and-drop) method. Ticks were determined by the species and all Ixodes ricinus were observed by a dark-field microscope. A total of 3,158 individual ticks were collected, out of which 2,797 (88,6%) were Ixodes ricinus, 175 (5,5%) Dermacentor reticulatus and 186 (5,9%) Rhipicephalus sanguineus. At forest sites there were collected 1,102 Ixodes ricinus, in park-forest regions 730 and in the parks 569. Infection rate of ticks by B. burgdorferi sensu lato at the forest sites ranged between 23,8 and 36,3%, in the park-forests region 23,2-32,6% and 23,8 to 40% in the parks. Twelve of the sites met the criteria for high potential risk and three of them for moderate risk. Except for Makis, at all the forest sites, the actual risk from March to April was certain, and in other months the risk varied. In the park-forest regions, the actual risk was registered 16 times, the possible risk 23 times, and the limited riosk once. At the sites belonging to the park areas, cetrain actual risk was established 3 times, possible risk 30 times and the limited 7 times At the control site Suplja Stena, the actual risk of transmission of B. burgdorferi sensu lato was registered during all the months of the reaserch. At the teritory of Belgrade there are favorable conditions for maintaining vector Lyme disease and the potential and actual risk of transmission of Borrelia burgdorferi sensu lato.

The Dobrava-Belgrade virus (DOBV) is a member of the Bunyaviridae family, genus Hantavirus, possessing a single-stranded RNA genome consisting of three segments, designated L (large), M (medium) and S (small). In this study, we present phylogenetic analysis of a newly detected DOBV strain isolated from Apodemus agrarius. Analysis was based on partial L and S segment sequences, in comparison to previously published DOBV sequences from Serbia and elsewhere. A phylogenetic tree based on partial S segment revealed local geographical clustering of DOBV sequences from Serbia, unrelated to host (rodent or human). The topology of the phylogenetic tree was confirmed with a high percent of completely or partially resolved quartets in likelihood-mapping analysis, whereas no evidence of possible recombination in the examined S segment data set was found.

Verotoxin-producing Esherichia coli (VTEC) is one of six pathogenicity groups of Eschericha coli. The reservoir for VTEC is the intestinal tract of domestic animals, primarily ruminants. Investigations in our geographic region have also demonstrated that domestic animals are a significant VTEC reservoir. In spite of this, sporadic diseases in humans caused by these agents are rare, and no epidemics have been registered so far. The question is, therefore, what percentage of VTEC isolated in our region have the characteristics of enterohaemorrhagic E. coli (EHEC) which cause intestinal diseases in humans. The aim of this study was to test the isolated bovine strains for some phenotypic and genotypic traits and to determine the percent of strains that belong to EHEC. A total of 105 VTEC strains isolated from cattle were tested for the presence of verotoxin (vtx) genes by PCR (polymerase chain reaction) method. All of them possessed one or more vtx genes. In accordance with our investigations, only four (3.8%) strains, of a total of 105 VTEC strains belong to groups of EHEC. It may be concluded that the majority of strains isolated from cattle in this part of the world do not have phenotypic traits typical for EHEC. Therefore human VTEC associated diseases in Serbia are rare, despite the fact that domestic animals frequently harbor VTEC.

Background/Aim. Despite numerous research of Lyme disease (LD), there are still many concerns about environmental of infectious agent of LD, as well as its prophylaxis, diagnosis and treatment. The aim of this work was to determine the risk of LD in relation to the way of removing ticks and duration of tick attachment. Methods. In the period from 2000 to 2007 a prospective study was conducted including persons with tick bite referred to the Institute of Epidemiology, Military Medical Academy, and followed for the occurrence of early Lyme disease up to six months after a tick bite. Epidemiological questionnaire was used to collect relevant information about the place and time of tick bites, the way of a removing tick, duration of tick attachment, remnants of a tick left in the skin (parts of the mouth device) and the signs of clinical manifestations of LD. Duration of tick attachment was determined on the basis of size of engorged tick and epidemiological data. Removed ticks were determined by the key of Pomerancev. Professional removing of attached tick was considered to be removing of tick with mechanical means by healthcare personnel. Fisher's exact test, Chi squares test and calculation of the relative risk (RR) were used for data analysis. Results. Of 3 126 patients with tick bite, clinical manifestations of LD were demonstrated in 19 (0.61%). In the group of subjects (n = 829) in which a tick was not removed professionally there were 17 (2.05%) cases with LD, while in the group of respondents (n=2 297) in who a tick was removed professionally there were 2 (0.09%) cases with LD after tick bite (RR, 23.55; p < 0.0001). The disease was most frequent in the group of respondents with incompletely and unprofessionally removed ticks (2.46%). In the groups of patients with unprofessionally but completely removed ticks LD occurred in 0.89%, while in the group of subjects with a tick removed by an expert, but incompletely in 0.78% cases. The disease occurred rarely in the group with a tick removed completely and professionally (0.05%). There was no case of LD in the group of patients with a tick removed within 24 hours. The longer time of exposure after 24 hours, the higher absolute risk of disease was reported. Conclusion. In prevention of Lyme disease it is important to urgent remove a tick, to use a correct procedure of removing and to remove the whole tick without any remnants.

Background/Aim. Lyme disease (LD) is a cosmopolitan disease from the group of zoonoses that in humans can affect skins, joints, heart and/or nervous system. Research conducted in endemic areas, shows not only that the population, which lives or works in tick habitats, is in a high risk for getting infected by LD cause, but also that every enviroment itself is specific in regard to both vectors and B. burgdorferi species. In our enviroment, research of seropositivity to B. burgdorferi in persons exposed to vectors of LD have not been conducted. The aim of this study was to determine whether there was a higher risk for getting B. burgdorferi infection in the fieldworks of JKP "Zelenilo Beograd" than in other persons. Methods. Risk for getting LD in the fieldworkers of JKP "Zelenilo Beograd" (park-maintanance company) was determined according to seropositivity, the number and infection rate of LD vectors, and entomological risk index (ERI) value. Serum samples of 34 fieldworkers of JKP "Zelenilo Beograd", and 35 persons of the control group, were tested for the presence of specific IgM and IgG anti-B. burgdorferi antibodies by using ELISA test. In the research localities, the number of ticks was estimated by using the flag hours method, and tick infection rate was determined by dark field microscopy, with magnification rate of 400×. The data were analyzed statistically using χ2 test, and Pearson's correlation analysis. Results. Out of 34 fieldworkes serum samples, 8 (23.5%) were positive on anti-B. burgdorferi antibody presence, while in the control group only one (2.9%) sample was positive (p < 0,05). Seropositivity rate varied from 0-66.6% among localities. Flag hours value among the localities ranged from 8.2-29.9, and tick infection rate ranged from 10.8- 22.3%. ERI value was 0.03-0.15. Correlation between flag hour rate and seropositivity, tick infection rate and seropositivity, and also ERI and seropositivity was established. Conclusion. Frequency of B. burgdorferi seropositivity was significantly higher in green-surface maintenance workers, than in the control group not exposed to these habitats. A significant correlation was found between anti-B. burgdorferi antibodies findings and the number of ticks, tick infection rate, and ERI. The results obtained in our study indicate that there was a significantly higher risk for getting LD cause in the fieldworkers than in the control group. .

Thiourea‐based iminophosphorane (BIMP) organocatalysts featuring SPhos‐ or BIDIME phosphine units have been developed and successfully applied in the asymmetric addition of nitromethane to N‐Boc‐protected trifluoromethyl aryl ketimines. α‐Trifluoromethyl β‐nitroamines were obtained in 40–82% isolated yields and 80–95% enantioselectivities. A careful evaluation of the catalytic activity of BIMPs indicates that the catalysts derived from the combination via Staudinger reaction of a chiral 1,2‐amino alcohol‐derived thiourea‐organoazide with electron‐rich phosphines, promote the aza‐Henry reaction on fluorinated ketimines with the highest enantioselectivity, leading to the amine featuring a tetrasubstituted stereocenter in up to 95% ee. The reaction was performed also on gram scale, without loss of enantioselectivity. [ABSTRACT FROM AUTHOR]

The use of stereoselective phase-transfer catalysis as a reliable method for the enantioselective synthesis of optically active α-amino acid derivatives using achiral Schiff base esters has been well-developed in batch in the last 40 years. Recently, continuous flow technology has become of great interest in the academy and industry, since it offers safer process operating conditions and higher efficiency compared to a traditional batch processing. Herein, we wish to report the first example of enantioselective phase transfer benzylation of alanine Schiff base ester, under continuous flow conditions. Two different methodologies were investigated: a liquid-solid phase transfer catalytic benzylation using a packed-bed reactor and a liquid-liquid phase transfer catalytic benzylation in continuous stirred-tank reactors. Liquid-liquid phase transfer process in flow showed slightly better productivity than the batch process, while solid-liquid phase transfer benzylation proved much more advantageous in terms of productivity and space-time yield. Furthermore, continuous flow system allowed the isolation of benzylated product without any work up, with a significant simplification of the process. In both cases, phase transfer asymmetric benzylation promoted by Maruoka catalyst demonstrated high enantioselectivity of target quaternary amino ester in flow, up to 93% ee. [ABSTRACT FROM AUTHOR]

Here, we report the design of a disposable single-drop voltammetric sensor for the quantitative determination of antipsychotic drug trifluoperazine (TFP). The sensor was built using inkjet-printed carbon nanotube (CNT) electrodes, which were modified with dimercaptosuccinic acid (DMSA)-coated magnetite nanoparticles uniformly dispersed over reduced graphene oxide nanosheets (DMSA/Fe3O4/RGO). The used modifying materials were characterized by electron microscopy techniques (transmission electron microscopy (TEM) and field emission-scanning electron microscopy (FE-SEM)), X-ray powder diffraction, ζ-potential measurements, dynamic light scattering (DLS), and electrochemical methods (cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS)). The developed sensor, best operated at pH 7 in the Britton–Robinson buffer solution (BRBS), shows linear electrocatalytic activity with TFP in the concentration range of 1–50 μM, a low detection limit of 0.54 μM, and excellent selectivity, repeatability, and reproducibility with an relative standard deviation (RSD) of 2.4%. A voltammetric approach using square wave voltammetry (SWV) is a sensitive technique under optimized conditions for the analytical determination of submicromolar amounts of TFP. Bare CNT and RGO- and DMSA/Fe3O4-modified CNT electrodes showed lower electrocatalytic activity than the DMSA/Fe3O4/RGO/CNT electrode. The development of this kind of TFP sensor based on nanoparticle-decorated graphene nanosheets can offer a tool for point-of-care applications as sensors in biomedicine. [ABSTRACT FROM AUTHOR]

Two non-steroidal antiinflammatory drugs indomethacin and mefenamic acid were coordinated to Ru(ii)–arenes to afford four new complexes. The cytotoxic activities of the ligands and ruthenium complexes were tested in three human cancer cell lines (K562, A549, MDA-MB-231) and non-tumour human fetal lung fibroblast cells (MRC-5) by MTT assay. Cytotoxicity studies revealed that indomethacin Ru(ii)–arene complexes 1 and 3 displayed good cytotoxicity and apparent cytoselective profiles. The IC50 values obtained in leukemia K562 cells were comparable to those of cisplatin (10.3 μM (CDDP), 11.9 μM (1) and 13.2 μM (3)). Flow cytometric analysis of 1 and 3 in triple-negative breast cancer MDA-MB-231 cells revealed an interesting mechanism of action. At IC50 concentrations, 1 and 3 arrested cell cycle progression in S phase and caused rapid accumulation of cells in sub-G1 phase (up to 48%), while Annexin V-FITC/PI staining showed simultaneous occurrence of apoptotic and necrotic cell populations at approximately similar levels of 20%. Measurement of reactive oxygen species (ROS) production by DCFH-DA staining confirmed the potential of 1 and 3 to increase ROS even more than cisplatin. The interaction of the complexes with serum albumins showed their potential ability to bind tightly and reversibly to albumins. The affinity of the complexes to calf-thymus DNA was investigated by UV-vis spectroscopy, viscosity measurements and fluorescence emission spectroscopy for competitive studies of the complexes with ethidium bromide, revealing that their interaction probably occurs via intercalation. Taken together, the results strongly suggest the potential of complexes 1 and 3 to alter cell cycle progression and cause DNA-damage by means of direct DNA-binding or indirectly by ROS production. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the presence of heavy metals/metalloids (Pb, Cd, Hg, Cu, Fe, Zn, As) in the muscle tissue of fish from the Danube River (two locations: Zemun and Grocka). For the purpose of heavy metal determination in fish muscle, 120 samples of six different fish species, Prussian carp, barbel, bream, carp, pike perch, and catfish were collected. For determining heavy metals, we used microwave oven digestion and atomic absorption spectrometer methods. The highest average content of Pb (0.084 ± 0.004 mg kg), Cd (0.082 ± 0.003 mg kg), Hg (0.466 ± 0.006 mg kg), and As (0.333 ± 0.007 mg kg) was found in the muscle of carp (an omnivorous fish) from Grocka, while the highest average level of Fe (13.60 ± 0.03 mg kg) was deposited in bream (also omnivorous) from Zemun. Also, the average Cu level (1.62 ± 0.13 mg kg) was the highest in catfish muscle (a carnivorous fish) from Grocka, while the highest Zn content (11.16 ± 0.17 mg kg) was determined in muscle of Prussian carp (an omnivorous fish) from Zemun. The highest content of heavy metals (Cu, Fe, and Zn, respectively) in muscle of the six different types of fish from both locations was symmetrically arranged by species (catfish, barbel, and Prussian carp, respectively). Concentrations of Pb, Hg, and As in the Danube River fish muscle were under the maximum residual levels prescribed by the European Union (EU) and the maximum allowed concentrations (MAC) for Serbia. On the other hand, in all fish muscle from both locations (Zemun and Grocka), higher concentrations of Cd than prescribed (MAC) were found, with the exception of bream and pike perch. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the influence of different feed composition and cultivation on the composition of fatty acids, their relationships, and the cholesterol content in the tissue of rainbow trout ( Oncorhynchus mykiss). Trout were reared in ponds at two different altitudes and raised on different feeds. The total amount of monounsaturated fatty acids in fillets of both groups showed no statistically significant difference. However, higher levels of n-6 fatty acids were found in Pond I trout fillets than in Pond II fillets, while Pond II fillets contained almost even amounts of n-3 and n-6 fatty acids. Total cholesterol content in Pond I trout fillets was significantly lower ( p < 0.05) than in Pond II fillets. The results indicate that the final composition of the trout fillets was mostly dependent on the feed the live fish had consumed; the altitude of the ponds had less effect. Also, both groups of trout could be used in the human diet as biologically valuable foods. Practical application: Our results show influence of altitude, feed, and fatty acid composition of feed on trout fillets. The average fatty acid level in the trout fillets increased in the order SFA

Two new platinum(IV) complexes (1, trifluoperazinehydrochloride-aquapentachloridoplatinate(IV) and2, chlorpromazine-chlorpromazinehydrochloridepentachloridoplatinate(IV)) were synthesized in the reaction of K2[PtCl6] with trifluoperazine dihydrochloride (TF·2HCl) or chlorpromazine hydrochloride (CP·HCl). The complexes were characterized by elemental analysis, molar conductivity measurement, and spectral (IR,1H,13C, 2D1H–13C heteronuclear correlation spectra,195Pt NMR, and MS) methods. Outer-coordination sphere was proposed for1; while in2, the ligand was coordinated to the metal. The complexes exhibit antibacterial effect on strains ofBacillus subtilis,Bacillus cereus,Bacillus pumilus, and methicillin-resistantStaphylococcias Gram-positive bacteria and anEscherichia colias Gram-negative bacteria, as well as the reference strains. [ABSTRACT FROM PUBLISHER]

The thermal decomposition of [RuCl(η- p-cymene)] ( 1) and its biologically active N-alkylphenothiazine compounds of composition L[RuCl(η- p-cymene)] where L = CPH ( 2), TFH·HCl ( 3), and TRH ( 4) (chlorpromazine hydrochloride, CP·HCl; trifluoperazine dihydrochloride, TF·2HCl; and thioridazine hydrochloride, TR·HCl, respectively) has been studied. The crystal and molecular structure of compound 3 was determined earlier by single crystal X-ray diffraction analysis. The thermal data were collected by simultaneous TG/DSC measurements. For evolved gas detection, the qualitative reaction of chlorides with AgNO in an acidic solution was applied. The measurements were carried out in the temperature range to 700 °C in nitrogen atmosphere. Compounds of L[RuCl(η- p-cymene)] crystallize with water or water/2-propanole. On the basis of thermal data, the trend in the solvent bonding energies was assessed. [ABSTRACT FROM AUTHOR]

OBJECTIVE: The aim of our study is to show the potential and actual risk of transmission of Borrelia burgdorferi at a number of localities in Belgrade. MATERIALS & METHODS: On the territory of Belgrade on 16 sites of which 15 were test and belonged to different environmental categories (5 parks, 5 park-forests and 5 forests), while 1 was control, we investigated potential and actual risk of transmission of B. burgdorferi. From March to October 2012, the ticks were collected once a month by using the fl ag-hours method. Ticks were determined by the species and all Ixodes ricinus were viewed by dark fi eld microscopy. RESULTS: A total of 3158 ticks were collected, of which 2797 (88.6%) Ixodes ricinus, 175 (5.5%) Dermacentor reticulatus and 186 (5.9%) Rhipicephalus sanguineus. In forest were collected 1102 I. ricinus, in park-forests 730 and 569 in parks. Twelve sites met the requirement of high potential risk and of moderate there were three. In all forest sites, the actual risk in March and April was not certain and in the other months it was variable. In park-forests certain actual risk was registered 16 times during investigation, possible risk 23 and limited just once. In parks, certain actual risk was established 3 times, possible risk 30 and limited 7 times. Of 16 sites, there was one control (not chemicaly treated) where actual risk was registered during all months for 3 times. CONCLUSION: In Belgrade there are favorable conditions for maintaining potential and actual risk of transmission B. burgdorferi. [ABSTRACT FROM AUTHOR]

By WHO defi nition, the term biosecurity refer to the protection of microbial agents from loss, theft, diversion or intentional misuse.Security is not a new concept in biological research and medical laboratories. Biosafety and biosecurity are related, but not identical concepts. In some cases, biosecurity practices may confl ict with biosafety practices. Designing a biosecurity program that does not jeopardize laboratory operations or interfere with the conduct of research requires a familiarity with microbiology and the materials that require protection. Protecting pathogens and other sensitive biological materials while preserving the free exchange of research materials and information may present signifi cant institutional challenges. A risk management methodology can be used to identify the need for a biosecurity program. The risks need to be identifi ed, prioritized and resources allocated based on that prioritization. Management, researches and laboratory supervisors must be committed to being responsible stewards of infectious agents and toxins. Development of a biosecurity program should a collaborative process involving all stakeholders. The stakeholders include but are not limited to: senior management; scientifi c staff; human resource staff; and safety, security and engineering offi cials. The involvement of organizations and personnel responsible for a facility's overall security is critical because many potential biosecurity measures may already be in place as part of an existing safety or security program.This coordinated approach is critical in ensuring that the biosecurity program provides reasonable, timely and cost-effective solutions addressing the identifi ed security risks without unduly affecting the scientifi c or business enterprise or provision of clinical and diagnostic services. [ABSTRACT FROM AUTHOR]

In the past decades there is growing awareness of the threat of bioterorrism which is defined as: «hostile use of microbes for political, religious and other ideological or criminal motivation». The risk of this threat is certainly augmented by the propulsion of scientific disciplines such as molecular biology and genetics, biotechnology, bioinformatics, and nanotechnology. Results and knowledge gained in these fields are typical dual-use products: from one side they serve for welfare of humankind, whereas from the other side they may be misused for deliberate impair of man and animal health as well as for the destruction of plant. In neutralizing bioterroristic act a major role play microbiology laboratory which principal goal is rapid detection and identification of biological agents. The main characteristics that determine laboratory abilities in managing these activities are existing levels of biosafety and biosecurity. Biosafety is the application of knowledge, techniques and equipment to prevent personal, laboratory and environmental exposure to potentially infectious agents or biohazards, i.e. biosafety defines the containment conditions under which infectious agents can be safely manipulated. Biosecurity is the effective implementation of measures to control access to pathogens and prevent unauthorized access. Vital elements that confere relevant level of biosafety and biosecurity are: laboratory equipment, staff competence,existing legislation, instructions and guidlines. If the use of biological weapon is suspected, basic principle during specimen processing is that screening and other tests should be performed in bisafety level 3 (BSL-3) rooms and cabinets, whereas in the case of probable use of highly contagious and lethal agents, laboratory tests should be carried out in the highest biocontainment level, BSL-4. Today definitive results are obtained in a significantly shorter period of time due to the expansion of molecular genetic methods, first of all PCR. Numerous other sensitive and specific methods are used with increasing frequency: immunodiagnostic (HHA in magnetic field, ELISA, ECORIGEN, TRF-Victor, Luminex- FC), multiple tests with high throughput, (Nanogen, Combimatrix, Affymetrix Resequencing), robotics (many sensors in one chip, high throughput), and biological mass spectrometry. Rationale protocol for the detection and identification of biological agents is as follow: 1. part of specimen, after inactivation (sterilization), is examined by (electron) microscopy; (result is obtained after 1-2h, preliminary); 2. part of specimen is seeded in nutrient media (liquid, enriched); after that PCR and/or immunological tests are performed (result after 6-12h, preliminary); 3. part of specimen is subjected to standard methods of isolation and identification (result after 2-5 days, definitive); 4. additional tests in animal model are carried out if virulence properties or other traits of agent in question are investigated; 5. part of specimen is saved (frozed) for additional investigation, if needed. It may be concluded that microbiological investigation play a key role in defending against bioterroristic attack. In dealing with this problem attention should be focused on: specimens handling, screening tests, speed, sensitivity and specificity of applied methods, reliability of obtained results , and on working conditions above all, taking into account existing levels of biosafety and biosecurity. [ABSTRACT FROM AUTHOR]